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Synthesis and characterization of a fluorescence probe of the phase transition and dynamic properties of membranes

Biochemistry 22(25): 5714-5722

Synthesis and characterization of a fluorescence probe of the phase transition and dynamic properties of membranes

The synthesis and characterization of a new fluorescence probe whose emission spectra, anisotropies and wavelength-dependent decay times are highly sensitive to the phase state of phospholipid vesicles is described. This probe is 6-palmitoyl-2-[[2-(trimethylammonio)ethyl]methylamino]naphthalene chloride (Patman). The emission maximum of Patman shifts from 425 to 470 nm at the bilayer transition temperatures. The spectral properties of Patman reveal nanosecond time-dependent spectral shifts, which are the result of membrane relaxation around the excited state of Patman. The apparent fluorescence lifetimes of Patman are strongly dependent upon the emission wavelength, and the fluorescence phase and modulation data prove that the spectral shifts are due to an excited-state process, and not ground-state heterogeneity. As expected, the fluorescence anisotropies reflect the phase transitions of the bilayers. In addition, the anisotropies are dependent upon the emission wavelength because the duration of the excited state varies across the emission spectrum. The different apparent lifetimes across the emission spectrum allow the relaxed and unrelaxed emission spectra to be resolved by phase-sensitive detection of fluorescence. Also, the emission spectra of Patman show marked shifts to longer wavelengths as the excitation wavelength is increased. These red-edge excitation shifts are sensitive to the temperature and phase state of the bilayers.

Accession: 006561697

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